ES2364419B1 - PHARMACEUTICAL COMPOSITIONS THAT INCLUDE AN INCLUSION COMPLEX FORMED BY DISULFIRAM AND A CYCLODEXTRINE, USEFUL IN THE TREATMENT OF THE ALCOHOL AND COCAINE DEPENDENCE. - Google Patents

Abstract

Pharmaceutical compositions comprising an inclusion complex formed by Disulfiram and a cyclodextrin useful in the treatment of alcohol and cocaine dependence.

Description

Pharmaceutical compositions comprising an inclusion complex formed by Disul fi ram and a cyclodextrin, useful in the treatment of alcohol and cocaine dependence.

The present invention relates to a new pharmaceutical composition comprising a disulphragm complex (DSF) with a cyclodextrin (CD), its method of production, and its use in the treatment of alcohol and cocaine dependence.

State of the art

Alcohol dependence is a multicausal pathology, so it must be addressed with a biopsychosocial treatment, which includes pharmacological support. There are several types of treatments for alcohol dependence, in which various pharmacological and / or psychotherapeutic tools are used; however, they have in common the inconvenience of presenting low adherence, which prevents the rehabilitation of the patient and their subsequent reintegration into society.

One of the most commonly used drugs in the treatment of alcohol dependence, is Disul fi ram, (1- (diethylthiocarbamoyldisulfanyl) -N, N-diethyl-methanethioamide; CAS No. 97-77-8), molecular weight of 296,543 g / mol , molecular formula C10H20N2S4, whose chemical structure corresponds to Formula (I):

The first report on DSF and its possible applications in the treatment of alcohol dependence appears in 1948, by Jacobsen and Hald (Jacobsen Erik Hald J., Larsen V. The sensitizing effect of tetraethyltiuram dysulphide (Antabuse) to ethyl alcohol, Acta Pharmacol et Toxicol, 4: 285-296, 1948). For years, this drug has been widely used as an ethyl alcohol rejection therapy.

DSF produces hypersensitivity to alcohol by inhibiting the enzymatic mechanism that oxidizes acetaldehyde to acetic acid, which occurs in the liver during normal ethanol catabolism. The absorbed ethyl alcohol is removed from the organism primarily (> 90%) through oxidative mechanisms located mostly in the liver, and mainly involving the enzyme alcohol dehydrogenase and to a lesser extent certain microsomal enzymes (microsomal oxidative system of alcohol). The first step in alcohol metabolization is to oxidize the substrate to acetaldehyde through the alcohol dehydrogenase (ADH) enzyme system that uses nicotinamide adenine dinucleotide (NAD +) as a hydrogen acceptor. The majority of acetaldehyde continues to oxidize acetic acid in the liver, due to a family of isoforms of the enzyme aldehyde dehydrogenase (ALDH), which also uses NAD + as a hydrogen acceptor (Hardman, Joel G .; Limbird, Lee E. Goodman & Gilman, The Pharmacological Bases of Therapeutics, 9th edition, Vol. I, McGraw-Hill Interamericana Editores SA, Mexico,

p. 417-418, 1996). Normally the ability to remove acetaldehyde by ALDH exceeds the ability to generate acetaldehyde by oxidation of ethanol, so that circulating levels of acetaldehyde are quite low.

DSF competes with NAD + for the enzyme ALDH producing an apparent irreversible inhibition of enzyme activity (McEvoy, Gerald. AHFS Drug Information, 44th edition, American Society of Health-System Pharmacists Inc., Bethesda, USA 3630, 2002). There is then an accumulation of acetaldehyde in the blood, which is responsible for the signs and symptoms, which have been referred to as "acetaldehyde syndrome" (ethanoldisul fi ram reaction, RED), which appears within 10 minutes after the intake of ethanol and which is mainly characterized by redness of the face, pulsatile headache, dyspnea, nausea, vomiting, perspiration, hypotension, chest pain, dizziness and blurred vision. The duration of the syndrome varies between 30 minutes and a few hours; subsequently there is tiredness and sleep. This syndrome can be triggered even by 7 mL of ethanol in sensitive subjects. People under treatment with DSF should be prevented in a timely manner that the use of shaving lotions or frictions, medications such as cough syrups or even sauces or fermented vinegar, can trigger the syndrome. The ethanol-disulphram reaction can occur up to 1-2 weeks after the last dose of the medication.

The active metabolites of DSF that cause clinically important effects in the body are: diethylthiocarbamate (DDC) and methyl diethylthiocarbamate (MeDDC).

DDC, in addition to inhibiting ALDH, inhibits dopamino-β-hydroxylase causing a decrease in norepinephrine and epinephrine contained in the heart, blood vessels and adrenal medulla, which would explain the hypotensive response and other cardiovascular reflexes of DSF (Nakako G, Holloway JE , Schackford, JS. Effects of disul fi nance on the cardiovascular responses to ethanolin dogs and guinea pigs. Toxicology and applied pharmacology, 14 (3): 439-446, 1969).

In the pharmaceutical market there are two pharmaceutical forms that correspond to pellets and oral tablets, the latter being the most used in the therapy of alcohol dependence.

The success of the use of oral DSF in the treatment of alcohol dependence depends on the patient's adherence to therapy for the appropriate period. This is explained because the patient simply stops taking DSF and / or cheats the therapist causing the treatment to be abandoned. All DSF therapy requires a support network composed of family and friends (O'Farrel TJ, Bayog RD. Antabuse contracts for married alcoholics and their spouses: A method to maintain Antabuse ingestion and decrease with fl icts about drinking. J Subst Abuse Treat: 3 : 1-8, 1986).

Benefit from the use of DSF has also been found in patients with cocaine dependence, with or without comorbid alcohol dependence (Carroll, KM, Ziedonis, D., O'Malley, SS, McCance-Katz, E., Gordon, L. and Rounsaville, BJ, Pharmacologic interventions for abusers of alcohol and cocaine: a pilot study of disulphram versus naltrexone, American Journal on Addictions 2, 77-19, 1993; Carroll K. et al. Efficacy of disulphilia and cognitive behavior therapy in cocaine- dependent outpatients Arch Gen Psychiatry, 61: 264-72, 2004. Preti A. New developments in the pharmacotherapy of cocaine abuse Addict Biol, 12 (2): 133-51, 2007; and Kenna GA, Nielsen DM, Mello P , Schiesl A, Swift RM, Pharmacotherapy of dual substance abuse and dependence, CNS Drugs, 21 (3): 213-37, 2007. Review).

It has been previously discussed that oral disulphram therapy has the disadvantage of the low rate of adherence to treatment and the patient simply stops taking the drug. A radical strategy to avoid abandonment of therapy was devised by Marie, who first introduced the subcutaneous implant of SDF (Marie, C. Propos d'un Nouveau Mode de Traitement de l'Alcoolisme Chronique par Implantation de Disulfure de Tetraethyle-Thiourane Thèse, Faculté de Médicine de l'Université de Paris, 1955). These implants or pellets have been used to improve the therapeutic use of disulphram, however, since the necessary therapeutic doses by mouth are high (over 200 mg / day), it has been suggested that these pellets, usually of they would reach effective pharmacological levels and that their effectiveness would be mainly placebo type (Wilson A., Davidson W., Blanchard R. Disul fi x implantation: A trial using placebo implants and two types of controls. J. Stud Alc, 41: 429-436, 1980 ).

Animal studies have shown that DSF can be effectively mobilized from the subcutaneous injection site if oil bioavailability is modified (Stromme JH .; Eldjarn L., Stensrud T., Distribution and chemical forms of diethyldithiocarbamate and tetraethyliuram disul fi de (disul fi ram) in mice in relation to radioprotection, Biochem Pharmacol 15: 287-297, 1966; and Phillips M., Gresser JG., Sustained-release characteristics of a new implantable formulation of disulphram. J. Pharm Sci 73 (12), 1718-1720 , 1984), dissolved in polyethylene glycol (Phillips,

M. Excretion of Carbon Disul fi de in Breath Following Subcutaneous Injection of Disul fi ram, Clinical Research, 1980; 28 (3), 623 A) or suspended in water (Phillips, Michael; Gresser, Joseph G. Sustained-Release Characetristics of a New Implantable Formulation of Disu fl iram, J. Pharm. Sci. Subst. Abuse, 1984; 73 (12) , 1718-1720). However, Phillips and Greenberg, (Phillips, M .; Greenberg, J. Dose-Ranging Study of Depot Disul fi ce in Alcohol Abusers, Alcohol Cli. Exp-Res. 1992; 16 (5): 964-967) demonstrated that the suspension in DSF water it would not act as a form of deposit.

Another modality of implant has been devised by Chandia et al., (Chandia R., Cid E., Vallejos C., Avilas JM. Clinical study of disulphram in injectable suspension. A new alternative in the treatment of alcoholism. Rev. Med. Chile, 118, 855-861, 1990), who inject the DSF into microgranules, suspended in a vehicle composed of propylene glycol / water, subcutaneously. The administration of 1 g by this system produced intense local pain and an inflammatory reaction although maintaining levels of the active metabolite of disulphram, diethyldithiocarbamate, higher than those of the traditional implant, for at least one month (Cid E. Pharmacokinetic study of disulphram administered in the form of an injectable suspension, thesis for the title of Chemist-Pharmacist of the University of Chile, 1986).

Phillips and Greenberg, (Phillips M .; Greenberg J. Dose-Ranking study of depot disulphilia in alcohol abusers. Alcohol Clin Exp Res. Vol 16 (5), 964-967, 1992) conducted a clinical study with two pharmaceutical formulations of DSF Deposit In the first formulation, from 1 to 3.5 g of DSF suspended in 5% carboxymethyl cellulose was administered, which resulted in a difficult injection due to the high viscosity, also presenting a low or no ethanol-disulphir reaction and irritation and pain local, which demonstrated the clinical ineffectiveness of this formulation. In the second formulation, the viscosity was reduced by administering from 60 to 90 mg / kg of DSF in 0.1% Tween 80, which allowed to facilitate administration and reduce local discomfort somewhat. However, this formulation had to be administered with an attack dose of 15 mg / kg of oral DSF to obtain the expected clinical efficacy.

The inventors have surprisingly found that pharmaceutical formulations comprising a DSF Inclusion Complex with a CD solve many of the difficulties tending to obtain effective treatments.

CN 1376463 A describes a pharmaceutical composition for ocular topical use comprising a CD and DSF, where the DSF has an improved solubility. The DSF and the CD are mixed in water and lyophilized. The final product is obtained by formulating a lyophilized powder with water ready for application to the eyes. This composition corresponds to a eye drops for the treatment of cataracts and is intended to increase the solubility of DSF, since the water solubility of this compound is a physicochemical limitation for different types of formulations, being around 0.02 mg / mL.

Wang S et al. (Wang S, Li D, Ito Y, Nabekura T, Bioavailability and anticataract effects of a topical ocular drug delivery system containing disid fi ram and hydroxypropyl-beta-cyclodextrin on selenite-treated rats. Current Eye Research. Vol. 29 (1), 51-58, 2004) conducted a study of eye drops containing a high concentration of DSF and a CD. The composition is oriented to the treatment of cataracts, and a high concentration of CD is used as a solubilizer to increase the solubility of DSF and obtain an appropriate pharmaceutical form to be applied directly to the eyes. In this document a physical mixture of DSF with CD was used, not demonstrating the formation of a complex but only the increase in solubility provided by the CD to the active substance.

EP 1574221 A describes injectable compositions of diclofenac complexed with a CD. This document does not suggest sustained release compositions of the active ingredient, but rather the CD is used to increase the solubility of the active substance. The use of CD in the formulation of this document gives stability to the parenteral product.

According to the background delivered through the state of the art, it is clearly visualized that it has not been completely successful to obtain an effective treatment for alcohol dependence. The inventors of the present invention have developed new formulations that allow, on the one hand, a good adherence to the treatment, which translates into a lower abandonment of therapy, and on the other, it has been possible to avoid the inconveniences of causative parenteral or subcutaneous administrations of pain and swelling at the site of inoculation. Thus, with the modified release pharmaceutical forms developed in the present invention, adequate therapeutic levels are achieved, with excellent results in terms of their bioavailability and most importantly with high clinical efficacy.

Therefore, an object of the present invention is to provide modified release dosage forms of DSF directed to solve the limitations of the prior art mentioned above and which can be administered orally, intravenously, intramuscularly and intradermally, among others.

Detailed description of the invention

The present invention describes new pharmaceutical compositions comprising a CD inclusion complex with DSF as active ingredient. In the present invention, hydroxypropyl-β-cyclodextrin (HP-β-CD) is used as an excipient for the formation of a complex with DSF (hereinafter "DSF-CD Complex"), which causes changes in the physicochemical properties of the drug . This association allows the formulation of pharmaceutical forms of DSF that contribute to solve the current problems of alcohol dependence treatment.

The inventors have surprisingly found that pharmaceutical compositions comprising the association of DSF and HP-β-CD in the form of an inclusion complex, improve clinical effectiveness by significantly increasing adherence to treatment.

On the other hand, the inventors have surprisingly also found that the association of DSF and HP-β-CD in the form of an inclusion complex formulated in the presence of uncomplexed DSF (hereinafter "free DSF") also improves clinical effectiveness, increasing Significantly adherence to treatment.

It is important to define here that in the context of the present invention "Physical Blending" means a mixture, in solid state, of clearly defined proportions of CD and DSF; and by "Binary Mix" to a mixture, in different proportions of free DSF and inclusion complex formed by the CD and DSF (DSF-CD Complex). The Binary Mix will be defined according to the proportions of free DSF with respect to DSF-CD Complex. Therefore, a 40:60 Binary Mix means that it contains a ratio of free DSF to DSF-CD Complex equal to

40:60

The results obtained with the parenteral formulations surprisingly point to an extended release formulation even in the presence of CD, a recognized solubilizing agent.

In order to provide a greater background with respect to inclusion complexes, these are defined as a form of chemical complex in which a molecule or part thereof, the host (in this case the active agent DSF), is encapsulated or included within of another molecule or structure made of said molecules, the host (in this case the CD). The inclusion complex with CD is then made up of host (DSF) and host (CD, in this case specific HP-β-CD). This association is characterized by the presence of weak intermolecular interactions, commonly called hydrophobic junctions similar to those found in various biological systems (enzyme-substrate, antigen-antibody, for example). One or more CD molecules may contain one or more host molecules.

The proportion surprisingly found by the inventors to form a DSF inclusion complex with CD for use in a modified release pharmaceutical formulation is DSF: CD from 1: 1 to 1: 2.

In the context of the present invention, "modified release" means: (i) in the case of oral administration when at least the absorption rate, the maximum plasma concentration (Cm'ax) and / or bioavailability ( ABC), are greater than with the free DSF; and (ii) in the case of parenteral administration, when at least the plasma concentrations and / or the duration of these are greater than those obtained with the free DSF.

The interactions involved in the inclusion process are weak and are attributed to van der Waals forces, London dispersal forces, dipole-dipole interactions and hydrophobic interactions. In aqueous solution, water molecules are normally found within the internal cavity of the cyclodextrins (polar-apolar interaction); However, since they are structures of a polar nature, their presence is not thermodynamically favorable. When a host is introduced into the CD, it displaces these water molecules. Both the host and the water molecules are thermodynamically favored. The result of the complexation is then a thermodynamically more favorable system: the repulsive forces between high enthalpy water molecules within the CD and the repulsive forces between the apolar host within the polar solution next to the interactions of the weak type already mentioned, they favor the formation of the inclusion complex (Connors, K. The Stability of Cyclodextrin Complexes in Solution, Chemical Reviews, 97 (5): 1325-1357, 1997; Szejtli, J., Introduction and General Overview of Cyclodextrin Chemistry, Chemical Reviews, 98 (5): 1743-1753, 1998; Duchêne, D., New Trends in Cyclodextrins and Derivates, Editions de la Santé, Paris, 1992).

Over time, cyclodextrins have been chemically modified in order to improve the physical-chemical properties of the original compounds. This is how today we have the HP-β-CD, which arises from subsequent alkylation reactions of β-cyclodextrin, substitutions that are originally introduced in order to improve the aqueous solubility.

The pharmacology of HP-β-CD has been extensively studied by various authors. It should be kept in mind that it is a synthetic carbohydrate, not subject to active transport and that it cannot cross lipid barriers such as cell membranes (due to its hydrophilicity and size); and therefore, its distribution depends on the route of entry to the organism.

The HP-β-CD is presented to the amorphous state, has excellent solubility, excellent stability, low tendency to crystallize and its safety and effectiveness have been established. Excellent tolerance for various routes of administration including intravenous administration has been demonstrated, it is not irritating to skin or eyes, and since HP-β-CD is not toxic through the parenteral route, it is a material compatible with the pharmaceutical design.

The absolute bioavailability is dose-dependent and less than 0.5%; It has a limited distribution and renal clearance corresponds to 80-90% of the total clearance.

The diversity of results regarding the pharmacokinetics of DSF after parenteral administration, is mainly due to the insolubility of the drug and the impossibility of having a suspension that facilitates its administration, solubility and modified release, with minimal impact on the area of application. For this reason, the use of cyclodextrins was considered due to their ability to interact with other molecules and form inclusion compounds, which can modify and confer certain drugs on a variety of characteristics that allow their use to be improved with maximum safety ( Szejtli J. Cyclodextrin News, Vol 9: 159,1995).

The present invention aims to solve the problems posed in the previous paragraphs related to the physical-chemical properties, pharmacokinetic limitations, administration and non-compliance of the DSF therapy, by designing a new pharmaceutical release formulation modified with therapeutic effect and causing minimum damage to the inoculation point (in the case of IM or SC administration), which comprises a DSF complex with a CD (DSF-CD Complex).

The DSF-CD Complex used in the present invention has been developed by the inventors at the University of Concepción, Chile, and corresponds to an Inclusion Complex characterized by its X-ray spectra, Differential Scanning Calorimetry (DSC), Electron Microscopy of Scanning (MEB), Nuclear Magnetic Resonance (NMR) and Infrared Spectroscopy (IR).

In order to solve the problem of the technique described here, the inventors not only developed the DSF-CD complex, but also had to investigate numerous formulation alternatives with various vehicles and excipients and in different routes of administration to finally obtain the appropriate pharmaceutical compositions that They manage to solve the problem of technique.

In a preferred embodiment, pharmaceutical compositions comprising the Binary Mixture (which as noted above corresponds to the mixture of DSF-CD Complex with free DSF) are disclosed in a liquid pharmaceutical form, where the solvent is aqueous or oily.

In another preferred embodiment, pharmaceutical compositions comprising the Binary Mixture in a solid pharmaceutical form are disclosed.

In another preferred embodiment, the possible pharmaceutical compositions for the administration of the DSF-CD Complex are established within which oral, buccal, intramuscular, subcutaneous, and dermal administration are included.

In a specific embodiment a form of parenteral administration of modified release of a composition comprising the Binary Mixture and the appropriate vehicles for formulating the compositions is described.

In another specific embodiment a form of oral administration (tablets) of modified release of a composition comprising the Binary Mixture and the appropriate vehicles for formulating the compositions is described.

In another specific embodiment, a modified subcutaneous solid delivery form (pellets) of a composition comprising the DSF-CD Complex and the appropriate vehicles for formulating the compositions is described.

In preferred embodiments of the present invention, HP-β-CD is used.

In pharmaceutical compositions comprising the DSF-CD Complex different vehicles can be used for a suitable suspension of the complex. These excipients are commonly used in the state of the art, and are selected from: water, glycerin, lecithin, polyethylene glycol 300, propylene glycol, miglyol, ethyl oleate, microcrystalline cellulose, magnesium stearate, Tween and mixtures thereof.

Among the excipients used for liquid pharmaceutical forms can be mentioned: miglyol, ethyl oleate, water, polyethylene glycol and propylene glycol.

Miglyol is a medium chain triglyceride extracted as an endosperm oil from coconut nucifera L. It is used in different pharmaceutical formulations including oral, topical and parenteral preparations. In parenteral formulations it is used for the preparation of intravenous emulsions, solutions and suspensions, being mainly investigated in total parenteral nutrition (TPN) in combination with long chain triglycerides.

Ethyl oleate is mainly used as a vehicle in parenteral preparations for intramuscular and subcutaneous use. Its characteristics are similar to almond oil and coconut oil, however it has the advantage of being less viscous than the oils mentioned above and is absorbed more quickly by the body's fine tissues.

The inventors have surprisingly found that a formulation containing the Binary Mixture in an aqueous vehicle has a prolonged clinical effect over time, so it is only necessary to administer once a week, or once every fifteen days or once a month, depending on the dose used.

Description of the fi gures

Figure 1: Comparative dissolution kinetics of different tablet formulations containing the Binary Mixture.

Figure 2: Comparative dissolution kinetics of the tablets containing the Binary Mixture, corresponding to a formulation of the present invention (Example 10), compared to the commercially available Antabus® tablets.

Figures 3 to 10: Histological sections of rat skin of the subcutaneous administration (SC) areas of 15 mg / Kg of DSF in the form of free DSF in physiological serum (SF) (A); DSF-CD complex in SF (B); Binary Mix 25:75 in SF (C); and control with SF (D). The histological sections were obtained at different post-administration times of each of the trials. Figure 3 corresponds to 8 hours post-administration; Figure 4, 24 hours after administration; Figure 5, 48 hours post-administration; Figure 6, at 72 hours post-administration; Figure 7, 96 hours post administration; Figure 8, at 120 hours post-administration; Figure 9, at 144 hours post administration and Figure 10 corresponds to 168 hours post administration.

Figure 11: Histological sections of rat muscle from the intramuscular administration (IM) areas, obtained at 24 and 168 hours post administration of 1.5 mg / Kg of miglyol (Control).

Figure 12: Histological sections of rat muscle from the IM administration areas of 15 mg / kg of DSF in the form of Binary Mix 40:60 in miglyol (A) and DSF in the form of Binary Mix 40:60 in oleate ethyl (B). Histological sections were obtained 24 hours after administration.

Figures 13, 14, 16-18: Histological sections of rat muscle from IM administration areas of 15 mg / Kg of DSF in the form of Binary Mixture 40:60 in miglyol (A); DSF in the form of Binary Mix 30:70 in miglyol (B); DSF in the form of Binary Mixture 40:60 in ethyl oleate (C).

The histological sections were obtained at different post-administration times of each of the trials. Figure 13 corresponds to 48 hours post-administration; Figure 14, at 72 hours post-administration; Figure 16, at 120 hours post-administration; Figure 17, at 144 hours post-administration; Figure 18, at 168 hours post administration.

Figure 15: Histological sections in rat muscle in IM administration areas of 15 mg / Kg of DSF in the form of Binary Mixture 40:60 in miglyol (A) and DSF in the form of Binary Mixture 40:60 in ethyl oleate (B). Histological sections were obtained at 96 hours post-administration.

Figure 19: Effect of subcutaneous administration in rats of 15 mg / Kg of DSF in the form of Binary Mixture

25:75 in water (A); in miglyol (B); and (C) DSF free in water, over the plasma concentrations of total DDC over time.

Figure 20: Effect of intramuscular administration in rats of 15 mg / Kg of DSF in the form of Binary Mixture

30:70 in miglyol (A); to the form of Binary Mix 40:60 in miglyol (B); to the 40:60 Binary Mixture form in ethyl oleate (C), over the plasma concentrations of total DDC over time.

Figure 21: Effect of oral administration of 500 mg of DSF in the form of Binary Mixture 30:70 (A) and Antabus® (B), on plasma concentrations of total DDC in healthy volunteers.

Examples of pharmaceutical compositions with the complex

A.-Examples of Pharmaceutical Compositions containing the DSF-CD Complex in liquid vehicles for parenteral formulations (suspensions)

Example 1

Binary Mix 30:70

Complex DSF-CD ................................ 2.54g (Equivalent to 450mg of DSF) DSFibre ................................................. ......... 1,050g Lecithin ................................................. .......... 0.069g Propylparaben ................................................. ...... 0.01% Methylparaben ................................................. ....... 0.1% Buffercitrate vehicle ................................................. .. 3mL

The pharmaceutical compositions of the DSF-CD Complex in liquid vehicles for parenteral formulations were prepared according to known techniques. The preparation protocol for Example 1, which is homologated to procedures for preparing similar compositions mentioned below, is described below.

The preparation of the formulation of Example 1 was carried out in a sterile environment.

In a mortar, DSF and lecithin were incorporated. They were macerated until a uniform mixture of both compounds was obtained. Propylparaben and methylparaben were solubilized in a first part of the vehicle. The DSF-CD complex was then added with the remaining vehicle and mixed until a lump-free paste was obtained. Finally, the suspension was packaged in previously sterilized glass ampoules.

Example 2

Binary Mix 40:60

DSF-CD Complex ............................................... ..... 2,66g DSFibre ................................................. ......... 0.53g Miglyolc.sp ............................................... ......... 8.87g

Example 3

Binary Mix 30:70

DSF Complex: CD ............................................... ..... 1.94g DSFibre ................................................. ......... 0.65g Miglyolc.sp ............................................... ......... 8.27g

Example 4

Binary Mix 40:60

DSF-CD Complex ............................................... ..... 2,66g DSFibre ................................................. ......... 0.53g Oleatodeetiloc.sp ............................................... ..... 8.85g

B.-Examples of Pharmaceutical Compositions containing the DSF-CD Complex for Solid Formulations for oral administration

The pharmaceutical compositions of the DSF-CD Complex in solid vehicles for tablet formulations were prepared according to known techniques and with conventional excipients for such purposes.

Different tablets were made in which the ratio between DSF and DSF-CD Complex was modified until the optimum formulation was obtained. The following formulation corresponds to a general formulation, detailed in Table 1, in which the concentration ranges for the main constituents of tablets weighing between 600 and 800 mg are established.

TABLE 1

General formulation for solid oral administration formulations

Of all the formulations tested, 6 formulations were chosen to perform their dissolution profile, which correspond to Examples 5 to 10 detailed in Table 2. These compositions exhibited decay times compatible with oral administration.

TABLE 2

Examples 5-10 of tablets of the present invention (*)

(*) The amounts of all the ingredients of Examples 5 to 10 are expressed in mg.

The general protocol for preparing oral tablets is described below.

Tablet Processing Protocol

one.

Sift the DSF-CD Complex obtained at the University of Concepción, Chile;

2.

Preparation of the 20% polyvinylpyrrolidone binder solution in alcoholic solution;

3.

Moisten the DSF with the binder solution;

Four.

Granulation itself, passing the wetted mass through a mesh or sieve;

5.

Dry the granulate at 40 ° C for 24 hrs;

6.

Homogenization of the particle size of the granulate by passing it through a sieve (Size 195 μm);

7.

Mix with the rest of the excipients (disintegrate, diluent and finally the lubricant);

8.

Compression of the final mixture.

The tablets thus obtained were subjected to the basic controls of solid formulations such as weight, hardness, disintegration and evaluation of active ingredient.

The dissolution kinetics of the tablets of Examples 5 to 10 (Table 2) were performed according to the conditions indicated in USP XXIV, using a dissolving equipment Bahama St., Northridge equipped with 6 glasses, apparatus 2: paddle method, a a speed of 50 revolutions per minute (rpm) for 60 min. and using a volume of 900 mL of aqueous phase (water pH = 6.8 and 0.1 N HCl independently) at 37 ± 0.5 ° C.

Independent samples were taken during the period indicated above with a fixed and automatic sampling system with manual volume replacement.

Each formulation was incorporated on the dissolution medium under stirring and temperature of the established medium. 5 mL samples were taken at intervals of 1.3, 6.10.20, 30, 40, 50, 60 minutes with medium replacement.

6 dissolution profiles were made with a n = 3.

Figure 1 shows the dissolution kinetics of the 6 formulations that were tested and corresponding to tablets containing the Binary Mixture, according to the speci fi cations indicated in Table 2 (Examples 5 to 10). At 10 minutes all formulations presented more than 5% of dissolved DSF and more than 15% at 60 minutes, reaching approximately 25% for Example 10. Of all the formulations tested, Example 10 presented the best dissolution profile, so for the bioavailability studies this formulation was chosen.

As can be seen in Table 2, Examples 5 to 10 presented appropriate decay times for oral administration, as well as having a sufficient hardness that allows for proper handling during the entire process of packing, distribution, storage and administration of the tablets

The formulation of Example 10 was compared with the tablets currently available on the market corresponding to Antabus®.

Figure 2 shows a kinetics of comparative dissolution of the tablets of Example 10 with Antabus® tablets. It is observed that with Antabus® tablets only less than 5% of dissolved DSF is reached at 60 min; on the other hand, with the tablets of the present invention, up to 25% of DSF dissolved in this time is achieved; as also a smaller dispersion in the results is obtained. In addition, it can be visualized, comparing Figures 1 and 2, that all the formulations in Figure 1, which correspond to formulations of the present invention (Examples 5 to 10) have a better dissolution profile than Antabus® in Figure 2 .

That is, any formulation chosen from the present invention would be better than the existing one in the market (Antabus®).

C.-Examples of Pharmaceutical Compositions containing the DSF-CD Complex for Solid Formulations of subcutaneous administration (pellets)

This formulation is performed by direct compression of the DSF-CD complex for the manufacture of pellets, according to the conditions indicated in Example 11.

Example 11

Each pellet contains:

DSF-CD complex equivalent to 25 mg of DSF

Dimensions of the pellets:

Diameter = 3mm

Length = 7mm

Pharmacological evaluation of pharmaceutical compositions with the complex

1. Evaluation of the inflammatory intensity in the administration area: Histopathological study in rats

Histopathological evaluation was performed for subcutaneous (SC) and intramuscular (IM) injections of free DSF, to the DSF-CD Complex form and to the Binary Mix form in ratios of 25:75 and 40:60. Water, water with 15% glycerin, miglyol and ethyl oleate were used as excipients.

Subcutaneous Administration (SC)

15 mg / kg of free DSF, DSF-CD Complex, Binary Mixture 25:75 and physiological serum (SF) were administered in SC form.

The comparative histological study between physiological serum and DSF in its various forms of administration can be seen in Figures Nº3a10.

Figure 3 shows histological sections of rat skin obtained 8 hours post-administration of the different administrations:

A. Free DSF injection: severe hyperemia is observed in the deep vascular plexus with hypodermis hemorrhage;

B. DSF-CD Complex Injection: moderate hyperemia and severe superficial and deep vascular plexus are observed, respectively, with the presence of inflammatory cells;

C. Binary Mixture Injection: mild presence of inflammatory cells and hyperemia of the deep vascular plexus is observed;

D. SF injection: severe hyperemia is observed in both vascular plexuses.

Figure 4 shows histological sections of rat skin, obtained 24 hours post-administration of the different administrations:

A. Free DSF injection: severe hyperemia of both plexuses, abundant presence of inflammatory cells and hemorrhage in hypodermis are observed;

B. Injection of DSF-CD Complex, mild hyperemia of both plexuses, mild presence of inflammatory cells, very little edema and hemorrhage in hypodermis are observed;

C. Binary Mixture Injection: moderate hyperemia of the deep vascular plexus and moderate presence of inflammatory cells;

D. SF injection: there is slight destructuring of collagen fibers.

Figure 5 shows histological sections of rat skin, obtained 48 hours post-administration of the different administrations:

A. Free DSF injection: severe destructuring of the collagen fibers, deep vascular plexus hyperemia and moderate hypodermis hemorrhage;

B. DSF-CD Complex Injection: Moderate restructuring of collagen fibers, deep vascular plexus hyperemia and hypodermis hemorrhage;

C. Binary Blend Injection: moderate hyperemia of the deep vascular plexus and mild presence of inflammatory cells are observed.

D. SF injection: moderate presence of inflammatory cells and hyperemia of the deep vascular plexus is observed.

Figure 6 shows histological sections of rat skin, obtained 72 hours post-administration of the different administrations:

A. Free DSF injection: moderate hyperemia of both plexuses is observed, with moderate presence of inflammatory cells and hemorrhage in hypodermis;

B. Injection DSF-CD Complex: moderate hyperemia of the deep vascular plexus and severe presence of inflammatory cells;

C. Binary Blend Injection: moderate hyperemia of the deep vascular plexus and poor presence of inflammatory cells are observed;

D. SF injection: moderate hyperemia with mild presence of inflammatory cells is observed.

Figure 7 shows histological sections of rat skin, obtained 96 hours post-administration of the different administrations:

A. Free DSF injection: moderate hyperemia of the deep vascular plexus is observed, with hemorrhage in hypodermis and severe presence of inflammatory cells;

B. Injection DSF-CD Complex: moderate hyperemia of the deep vascular plexus is observed with mild presence of inflammatory cells, neutrophils in diapsis and edema in the dermis;

C. Binary Mixture Injection: moderate presence of inflammatory cells and hyperemia of the deep vascular plexus is observed. With mild bleeding in hypodermis;

D. SF injection: mild hyperemia of the deep vascular plexus is observed.

Figure 8 shows histological sections of rat skin, obtained 120 hours post-administration of the different administrations:

A. Free DSF injection: moderate hyperemia of the deep vascular plexus, moderate hyperemia of the deep vascular plexus, with a slight presence of inflammatory cells and collagen fibrosis derangement.

B. Injection DSF-CD Complex: mild hyperemia of the deep vascular plexus, presence of inflammatory cells and hemorrhage in hypodermis are observed;

C. Binary Blend Injection: mild hyperemia of the deep vascular plexus and poor presence of inflammatory cells are observed;

D. SF injection: mild hyperemia of the deep vascular plexus is observed, with little presence of fibroblasts.

Figure 9 shows histological sections of rat skin, obtained 144 hours post-administration of the different administrations:

A. Free DSF injection: moderate hyperemia of the deep vascular plexus and poor presence of inflammatory cells;

B. DSF-CD Complex Injection: low hyperemia of both plexuses and moderate presence of inflammatory cells is observed;

C. Binary Blend Injection: moderate hyperemia of the deep vascular plexus is observed with little presence of inflammatory cells;

D. SF injection: mild hyperemia of the deep vascular plexus is observed.

Figure 10 shows histological sections of rat skin, obtained 168 hours post-administration of the different administrations:

A. Free DSF injection: moderate hyperemia of the deep vascular plexus is observed;

B. DSF-CD Complex Injection: Severe deep vascular plexus hyperemia is observed with moderate hemorrhage in subcutis and abundant presence of phyblablasts;

C. Binary Mixture Injection: mild hyperemia of the deep vascular plexus is observed;

D. SF injection: hyperemia of the superficial and deep vascular plexus is observed.

When analyzing the results of histopathological lesions with the administration of free DSF versus the DSF-CD Complex and Disul fi rm to the Binary Mix form, it can be seen that the DSF produces a collagen fibrostructuring which progressed from mild, moderate to severe from 8, 24 and 48 hours post inoculation, damage that decreased to moderate in the case of the DSF to the complex form and to a mild character in the case of DSF to the Binary Mix form.

The presence of inflammatory cells, hyperemia of the superficial vascular plexus and hemorrhage in subcutis was moderate to severe in the controls between 24 and 96 hours in the case of free DSF, unlike what happened in the DSF to the complex form where the great presence of inflammatory cells occurred at the beginning in the control of 8 hours persisting with a moderate character until 72 hours. The hyperemia of the superficial vascular plexus was mild and of short duration the first 24 hours, while edema and hemorrhage in subcutis decreased to a moderate character in the case of the 24 and 72 hour controls. In the case of the DSF in the form of Binary Mixture, a significant decrease in the lesions was observed where there was a slight agglomeration of inflammatory cells and hemorrhage in the subcutis. The hyperemia of the deep vascular plexus in the case of free DSF was severe in the biopsies at 8 and 24 hours, remaining moderate to mild in subsequent controls, case by part was the cyclic behavior of this lesion in rats treated with DSF to the complex form, where the severity of the lesion was observed at the beginning and end of the study (8-168 hours), having a mild to moderate character in the intermediate controls. In the case of the DSF in the form of Binary Mixture, this lesion was observed in its mild to moderate character during all the controls of the study.

In conclusion, the SC administration of DSF produces severe histopathological lesions which are attenuated with the administration of DSF in the form of a complex or Binary Blend.

Intramuscular administration

Histopathological lesions were evaluated in intramuscular administration in rat muscle, obtained post-administration of 15 mg / Kg of DSF in the form of Binary Mixture in the proportions of 30:70 and 40:60 in 500 μL of miglyol or 500 μL of oleate ethyl.

Figure 11 shows a histological section in rat muscle, in areas of IM administration, obtained at 24 and 168 hours post-administration of 1.5 mg / kg of miglyol as a control.

A. 24 hrs. post-administration: mild hyperemia is observed in the perimysium, with diapédesis of neutrophils and lymphocytes, with little presence of inflammatory cells.

B. 168 hrs. post-administration, where normal muscle can be seen, only with a low hyperemia of the vessels located in the perimysium, with slight thickening of the collagen fibers in endomysium and perimysium.

Figure 12 shows a histological section obtained 24 hr post-administration:

A. Binary mixture (40:60) in miglyol: normal muscle fibers and scarce lymphoplasmocytic cells are observed, in epimisio;

B. Binary Mixture (40:60) in ethyl oleate: small areas of destructuring in muscle fibers are observed. Lymphoplasmocyte population in moderate quantity, in perimysium.

Figure 13 shows a histological section obtained 48 hours post-administration:

A. Binary mixture (40:60) in miglyol: moderate amount of inflammatory cells of lymphoplasmocytic type and poor alteration of muscle cells is observed;

B. Binary mixture (30:70) in miglyol: moderate amount of inflammatory cells of lymphoplasmocytic type is observed, in epimisio;

C. Binary Blend (40:60) in ethyl oleate: large areas of muscle fibrostructuring with loss of nuclei and rupture of fibers are observed. Abundant inflammatory population, in epimisio and perimisio. With areas of hemorrhage in perimysium.

Figure 14 shows a histological section obtained 72 hours post-administration:

A. Binary mixture (40:60) in miglyol: moderate to severe hyperemia is observed in vessels of the epimisio, with abundant amount of inflammatory cells, lymphoplasmocytic type and moderate to severe increase of fibroblasts and satellite cells.

B. Binary mixture (30:70) in miglyol: moderate to severe hyperemia is observed in epimisial vessels and moderate to abundant lymphoplasmocyte population, in epimisio;

C. Binary Blend (40:60) in ethyl oleate: Destructuring in muscle fibers, with loss of nuclei and rupture of fibers. Mild hyperemia in perimysium, with moderate in fl ammatory population, in perimysium and epimysium.

Figure 15 shows a histological section obtained 96 hours post-administration:

A. Binary Blend (40:60) in miglyol: it is observed with normal muscle fi bile structure and low to moderate inflammatory population, in epimisio;

B. Binary Mixture (40:60) in ethyl oleate: small areas of muscle fibrostructuring are observed, moderate presence of lymphoplasmocytic cells in epimisio, and moderate to severe hyperemia, in perimysium and epimisio.

Figure 16 shows a histological section obtained 120 hours post-administration:

A. Binary mixture (40:60) in miglyol: areas of fi bers with loss of nuclei are observed. Mild to moderate hyperemia. Abundant amount of lymphoplasmocytic inflammatory cells. Moderate increase in population of fibroblasts and satellite cells;

B. Binary mixture (30:70) in miglyol: normal muscle fibers and moderate lymphoplasmocytic presence are observed in epimisio;

C. Binary mixture (40:60) in ethyl oleate: lymphoplasmocytic inflammatory cells are observed in medium quantity in perimysium, with regular hypermia in the same area of connective tissue.

Figure 17 shows a histological section obtained 144 hours post-administration:

A. Binary mixture (40:60) in miglyol: slight alteration of muscle fibers is observed, with loss of nuclei. Mild hyperemia in perimysian vessels. Moderate amount of lymphoplasmocytic inflammatory cells;

B. Binary Mixture (30:70) in miglyol: small areas of muscle fibrostructuring are observed, with loss of nuclei, moderate lymphoplasmocyte population, in epimisio;

C. Binary mixture (40:60) in ethyl oleate, normal muscle fi les structure is observed, with few lymphoplasmocytic cells, in epimisio.

Figure 18 shows a histological section obtained 168 hours post-administration:

A. Binary mixture (40:60) in miglyol: there is a slight alteration of muscle fibers, with loss of nuclei and a small amount of lymphoplasmocytic inflammatory cells, in epimisio;

B. Binary Mixture (30:70) in miglyol: small areas of destructuring are seen in muscle fibers, with loss of nuclei. Moderate to abundant inflammatory population, in epimisio;

C. Binary mixture (40:60) in ethyl oleate: normal muscle fiber structure and a moderate to abundant inflammatory population are observed.

From the results obtained it is concluded that the Binary Mixtures are compatible with the IM administration route, since it produces minimal alterations in the tissue after its administration.

Pharmacokinetic Studies

In addition to the histopathological studies in rats, pharmacokinetic studies were performed in which the plasma concentrations of the DSF metabolites (DDC and MeDDC) were measured after SC (Figure 19) and / or IM (Figure 20) administration of 15 mg / kg DSF weight to the Binary Mix form.

The total plasma DDC concentration corresponding to the sum of the DDC and MeDDC metabolites was quantified, and was used as a parameter for the analysis of the results shown below.

Figure 19A shows the effect of SC administration of DSF in the form of Binary Mixture (25:75) using water as a solvent, on plasma concentrations of total DDC over time. The Cm´ax of 0.17 μg / mL was reached at the time of administration and detectable concentrations of total DDC were maintained even up to 168 hours post-administration, obtaining an area under the curve (ABC) of 3.31 ( μg xh) / mL.

Figure 19B corresponds to the effect of the SC administration of DSF in the form of a Binary Mixture (25:75) using miglyol as a solvent on the plasma concentrations of total DDC over time. The Cm´ax of 0.15 μg / mL was also reached at the time of administration and, as in the previous case, detectable concentrations of total DDC were maintained even up to 168 hours post-administration, obtaining an area under the curve (ABC) of 7.12 (μg xh) / mL.

Figure 19C corresponds to the effect of SC administration of free DSF using water as a solvent on the plasma concentrations of total DDC over time. The 0.25 μg / mL Cm´ax was also reached at the time of administration and significantly higher concentrations of total DDC were maintained during the first 144 hours post-administration, obtaining an area under the curve (ABC) of 4 , 86 (μg xh) / mL. At 168 hours post administration no plasma levels of active metabolites were detected.

Figure 19 shows that with the SC administration of the formulations containing the Binary Mixture (Figure 19A and 19B) better results were obtained, since although in all cases the bioavailability (ABC) did not significantly increase; detectable plasma concentrations of active metabolites (total DDC) were achieved for a longer time, with respect to SC administration of free DSF (Figure 19C); which suggests a longer therapeutic effect for the formulations of the invention. Even when a higher ABC was not reached since the formulation of Figure 19A is only a suspension of Binary Mixture in water, without some adjuvant that improves the characteristics of the formulation.

Figure 20A shows the effect of IM administration of the Binary Mixture (30:70) using miglyol as a solvent on plasma concentrations of total DDC over time. The Cm´ax of 0.17 μg / mL was reached at the time of administration, as it was when administered via SC (Figure 19). At 168 hours post-administration, detectable concentrations of total DDC were still observed, obtaining an area under the curve (ABC) of 6.05 (μg x h) / mL.

Like the previous Figure, Figure 20B shows the effect of IM administration of the Binary Mixture in miglyol but in a ratio of 40:60, on the plasma concentrations of total DDC over time. In this case, the Cm 'was 0.17 μg / mL at the time of administration and detectable levels of total DDC were maintained.

ax

until 168 hours post-administration, with an area under the curve (ABC) of 5.67 (μg x h) / mL.

Figure 20C shows the effect on plasma concentrations of total DDC, after IM administration of the Binary Mixture (40:60), using ethyl oleate as solvent. A smaller Cm´ax, of 0.10 μg / mL was reached 24 hours after administration. In addition, with this formulation in ethyl oleate, plasma levels of DDC were more erratic, since at 48 hours no active metabolites were detected in the plasma but at longer times if they were detectable, reaching an area under the curve (ABC ) greater than 7.06 (μg xh) / mL, compared with the same proportion of Binary Mixture but in miglyol as solvent (Figure 20B).

With all the proportions of Binary Mixtures and with the different vehicles used, with the IM administration, areas under the curve were obtained greater than with an SC administration. This shows that the IM pathway is clinically superior.

2. Clinical studies in humans

A. Bioavailability studies in healthy volunteers

A clinical protocol was performed for the study of bioavailability of DSF tablets in healthy volunteers, using the formulation of Example 10 of the present invention.

The Products investigated were:

Reference product: Antabus® 500 mg tablets. Paci fi c Farmochemical Laboratories (Formulation A).

Test product: 250 mg tablets. Faculty of Pharmacy, Universidad de Concepción, Chile; corresponding to Example 10 of the present invention (Formulation B).

Experimental design:

The design consists in studying the bioavailability of two formulations: a reference (Formulation A) and a product of the invention (Formulation B) in two periods and two sequences.

In the first period, half of the subjects received Formulation A (1 tablet of Antabus® 500 mg) in the second period they received Formulation B (2 tablets of the innovative formulation of 250 mg each). For the other half of the voluntary subjects the situation is exactly the opposite.

Subjects

The population of male subjects for Bioavailability studies should be as homogeneous as possible.

Number of subjects: 8 volunteers.

Analytical methodology:

An analytical method based on High Performance Liquid Chromatography (HPLC) with UV detection was used for the determination of volunteers' DDC and MeDDC in plasma, previously validated for pharmacokinetic studies.

The results of Figure 21 show that with the administration of the formulation of the invention corresponding to Example 10 (Figure 21A), areas under the curve (ABC) were obtained significantly larger than those obtained with the reference formulation (Antabus ®) (Figure 21B).

The ABC obtained with the tablets of the present invention is 0.617 (μg x h) / mL, while the same parameter for the tablets corresponding to the reference formulation was 0.227 (μg x h) / mL.

This indicates that with the formulation comprising the Binary Mixture, higher total DDC plasma levels are achieved than with Antabus® tablets at the same administered doses, which will allow the doses administered to be viable to achieve the same therapeutic effect.

B. Clinical effectiveness studies in patients with alcohol dependence

A liquid formulation for IM administration of the present invention (Example 1) was tested in patients of the Unit of Complex Drug Dependencies and Dual Pathology of the Psychiatry Service of Guillermo Grant Benavente Hospital, an establishment that meets the basic infrastructure and personnel requirements suitable for carry out phase II studies.

The study involved patients with alcohol dependence, who prior to the start of the study signed an Informed Consent. The alcohol dependent criteria were according to ICD-10, of the International Classification of Diseases, version number 10, of the World Health Organization.

The formulation of Example 1 of the present invention was administered IM to 29 patients in a dose of 1 g once a month.

Another 19 patients received the Antabus® reference formulation orally at a dose of 500 mg per day.

To determine the clinical effect achieved, the following were measured:

•

Withdrawal and Adherence Levels at 3 months of treatment;

•

Withdrawal and Adherence Levels after 6 months of treatment.

Parenteral administration was well tolerated and there was no reaction to a foreign body.

The results are given below in relation to the parameters measured in the treated patients: Abstinence and Adherence to therapy.

TABLE 3

Clinical effectiveness of an IM formulation according to the present invention and Antabus®, in the treatment for alcohol dependence

Abstinence is related to the number of days the patient does not consume alcohol.

Adherence corresponds to the actual number of therapeutic activities of patient participation regarding their initial commitment to treatment.

With the formulation of the invention a better clinical response is obtained, observing a greater Abstinence and Adherence, compared to conventional therapy (Antabus®).

These results confirm that with an IM administration once a month, better clinical results were achieved than with a daily oral administration of the tablets currently on the market.

Claims (15)

1. Pharmaceutical composition characterized in that it comprises an inclusion complex consisting of disulfiram (DSF) and cyclodextrin (CD), free or uncomplexed DSF and one or more pharmaceutically acceptable carriers. 2. Pharmaceutical composition according to claim 1 characterized in that the CD is hydroxypropyl-cyclodextrin. 3. Pharmaceutical composition according to claim 1 characterized in that the composition is in a solid pharmaceutical form. 4. Pharmaceutical composition according to claim 3 characterized in that the solid pharmaceutical form is oral or subcutaneous administration. 5. Pharmaceutical composition according to claim 1 characterized in that the composition is in a liquid pharmaceutical form. 6. Pharmaceutical composition according to claim 5 characterized in that the liquid pharmaceutical form is orally or parenterally administered. 7. Pharmaceutical composition according to claim 6 characterized in that the liquid pharmaceutical form is of subcutaneous or intramuscular administration. 8. Pharmaceutical composition according to claim 1 characterized in that the ratio of free DSF to DSF to the complex form is in the range of 20:80 to 40:60. 9. Pharmaceutical composition according to claim 8 characterized in that the ratio of free DSF to DSF to the complex form is 25:75. 10. Pharmaceutical composition according to claim 8 characterized in that the ratio of free DSF to DSF to the complex form is 30:70. 11. Pharmaceutical composition according to claim 8 characterized in that the ratio of free DSF to DSF to the complex form is 40:60. 12. Pharmaceutical composition according to claims 1 and 6, characterized in that the pharmaceutically acceptable carrier for liquid pharmaceutical forms is selected from water, miglyol and ethyl oleate, glycerin, lecithin, polyethylene glycol 300, propylene glycol, microcrystalline cellulose, magnesium stearate, Tween and mixtures thereof. 13. Use of the pharmaceutical composition according to claims 1 to 12, characterized in that it serves for the preparation of a medicament for the treatment of alcohol dependence. 14. Use of the pharmaceutical composition according to claims 1 to 12, characterized in that it is used for the preparation of a medicament for the treatment of cocaine dependence. 18 SPANISH OFFICE OF PATENTS AND BRANDS SPAIN REPORT ON THE STATE OF THE TECHNIQUE 51 Int. Cl.: See Additional Sheet twenty-one Application no .: 201090050 22 Date of submission of the application: 23.12.2008 32 Date of priority: RELEVANT DOCUMENTS

Category

Documents cited Claims Affected

X

NAGAI, N. et al .: "Delay in ICR / f rat lens opacification by the instillation of eye drops containing 1-15

disulfiram and hydroxypropyl-beta-cyclodextrin inclusion complex ". Biol. Pharm. Bull, 2007, vol. 30

(8), August, pages 1529-1534, the whole document.

X

WANG, S. et al .: "Bioavailability and anticataract effects of a topical ocular drug delivery system 1-15

containing disulfiram and hydroxypropyl-beta-cyclodextrin on slenite treated rats ". Current Eye

Research, 2004, vol. 29, (1), pages 51-58, the whole document.

TO

Treating alcoholic cocaine users with disulfiram and naltrexone. Pharmacology Biochemistry and 1-15

Behavior, 1992, vol. 42 (2), pages 366-367, page 367.

TO

PETRAKIS, I.L et al .: "Naltrexone and disulfiram in patients with alcohol dependence and comorbid 1-15

post-traumatic stress disorder ". Biol. Psychiatry, 2006, vol. 60, pages 777-783,

whole document.

Category of the documents cited X: of particular relevance Y: of particular relevance combined with other / s of the same category A: reflects the state of the art O: refers to unwritten disclosure P: published between the priority date and the date of priority submission of the application E: previous document, but published after the date of submission of the application

This report has been made • for all claims • for claims no:

Date of realization of the report 22.08.2011

Examiner H. Aylagas Cancio Page 1/4

TECHNICAL STATUS REPORT Application number: 201090050 CLASSIFICATION OBJECT OF THE APPLICATION A61K31 / 145 (2006.01) A61K47 / 40 (2006.01) A61P25 / 32 (2006.01) A61P25 / 36 (2006.01) Minimum documentation sought (classification system followed by classification symbols) A61K, A61P Electronic databases consulted during the search (name of the database and, if possible, search terms used) INVENES, EPODOC, WPI, EMBASE, BIOSIS, MEDLINE, NPL, XPESP, HCAPLUS. State of the Art Report Page 2/4  WRITTEN OPINION Application number: 201090050 Date of Written Opinion: 08.28.2011 Statement

Novelty (Art. 6.1 LP 11/1986)

Claims 3,5,7-15 Claims 1,2,4,6 IF NOT

Inventive activity (Art. 8.1 LP11 / 1986)

Claims Claims 1-15 IF NOT

It is considered that the application complies with the industrial application requirement. This requirement was evaluated during the formal and technical examination phase of the application (Article 31.2 Law 11/1986).  Opinion Base.- The present opinion has been made on the basis of the patent application as published. State of the Art Report Page 3/4  WRITTEN OPINION Application number: 201090050 1. Documents considered.- Below are the documents belonging to the state of the art taken into consideration for the realization of this opinion.

Document

Publication or Identification Number publication date

D01

NAGAI, N. et al.: "Delay i n ICR / f r at l ens opac ification by t he instillation of eye drops containing disulfiram and hydroxypropylbeta-cyclodextrin inclusion complex". Biol. Pharm. Bull, 2007, vol. 30 (8), August, pages 1529-1534, the whole document.

D02

WANG, S. et al.: "Bioavailability a nd ant icataract effects of a topical occu lar dr ug of the ivery system containing d isulfiram and hydroxypropyl-beta-cyclodextrin on slenite treated rats". Current Eye Research, 2004, vol. 29, (1), pages 51-58, the whole document.

D03

Treating alcoholic cocaine users with disulfiram and naltrexone. Pharmacology Biochemistry and B ehavior, 1992, vo l. 42 (2), pages 366-367, page 367.

D04

PETRAKIS, I.L et al .: "Naltrexone and disulfiram in patients with alcohol dependece and comorbid post-traumatic stress disorder". Biol. Psychiatry, 2006, vol. 60, pages 777-783, the whole document.

2. Statement motivated according to articles 29.6 and 29.7 of the Regulations for the execution of Law 11/1986, of March 20, on Patents on novelty and inventive activity; quotes and explanations in support of this statement The present application refers to a pharmaceutical composition characterized by comprising an inclusion complex consisting of disulfiram and cyclodextrin, optionally free or complexed sulfur and its use in the preparation of a medicament for the treatment of alcohol and cocaine dependence. Documents D1 and D2 refer to the use of eye drops that contain an inclusion complex formed by disulfiram and hydroxypropyl-beta-cyclodextrin in the treatment of cataracts. These documents cite the use of these inclusion complexes in order to increase the solubility of disulfiram in the eye drops and the permeability of the active substance in the eye. Therefore, in view of documents D1 and D2, the subject of claims 1, 2, 4 and 6 lacks novelty and inventive activity according to articles 6.1 and 8.1 of the L.P. Claims 3, 5, 7-15 lack inventive activity for the following reasons: Claims 3, 9-12 relate to the use of free or uncomplexed disulfiram in the pharmaceutical composition together with the inclusion complex formed by disulfiram and cyclodextrin. Already known are both compositions with a weapons activity that lead to the complex of exclusion (see documents D 1 or D 2 cited above) as pharmaceutical compositions that carry disulfiram alone (see documents D3 or D4), such a combination would be an option that an expert in the field could carry out with a reasonable expectation of success, therefore such claims lack inventive activity according to Article 8.1 of the LP Claims 14 and 15 refer to the use of the pharmaceutical composition according to claim 1 to prepare a medicament for treating alcohol and cocaine dependence. It is well known that disulfiram is an active substance used in the treatment of alcohol and cocaine dependence (see documents D3 or D4). Likewise, the use of cyclodextrins in the formation of complexes in order to increase the solubility of different active ingredients and among them disulfiram is also known (see documents cited above D1 or D2). Of course, it would be obvious even if it was a matter of experience in the matter that the completeness of the exclusion would also have the same pharmacological activity as the active principle that composes it, that is, its application in the treatment of alcohol dependence. and cocaine. Nor is it possible to economize preventive activity for claims 5, 7, 8 and 1 and to the various forms of administration by oral, parenteral, subcutaneous or intramuscular route are known administration forms in the state of the art. . Only the best clinical results with the intramuscular administration of the formulation of Example 1 seem proven in the description. Therefore, the matter corresponding to claims 3, 5, 7-15 lacks inventive activity according to Article 8.1 of the LP State of the Art Report Page 4/4